Bottom Line:
However, their combined role in cancer-associated deep vein thrombosis (DVT) and the molecular mechanisms, involved in its pathophysiology, needs further investigations.Significant correlation between data obtained in plasma and monocyte samples was observed.NF-kB inhibition was associated with decreased levels of all molecules in both cancer DVT+ and DVT-.

Affiliation: Department of Biomedical and Biotechnological Sciences, Section of General & Clinical Pathology and Oncology, University of Catania, Catania, Italy.

ABSTRACTSeveral studies highlight the role of inflammatory markers in thrombosis as well as in cancer. However, their combined role in cancer-associated deep vein thrombosis (DVT) and the molecular mechanisms, involved in its pathophysiology, needs further investigations. In the present study, C-reactive protein, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1β), matrix metalloproteases-9 (MMP-9), vascular endothelial growth factor (VEGF), tissue factor (TF), fibrinogen and soluble P-selectin, were analyzed in plasma and in monocyte samples from 385 cancer patients, of whom 64 were concomitantly affected by DVT (+). All these markers were higher in cancer patients DVT+ than in those DVT-. Accordingly, significantly higher NF-kB activity was observed in cancer patients DVT+ than DVT-. Significant correlation between data obtained in plasma and monocyte samples was observed. NF-kB inhibition was associated with decreased levels of all molecules in both cancer DVT+ and DVT-. To further demonstrate the involvement of NF-kB activation by the above mentioned molecules, we treated monocyte derived from healthy donors with a pool of sera from cancer patients with and without DVT. These set of experiments further suggest the significant role played by some molecules, regulated by NF-kB, and detected in cancer patients with DVT. Our data support the notion that NF-kB may be considered as a therapeutic target for cancer patients, especially those complicated by DVT. Treatment with NF-kB inhibitors may represent a possible strategy to prevent or reduce the risk of DVT in cancer patients.

pone.0132496.g001: Cytokines secretion in monocytes from cancer patients with and without DVT.Levels of IL-6, TNF-α, IL-1β, and VEGF were measured in supernatants of purified monocytes from cancer patients with and without DVT by a sensitive enzyme-linked immunosorbent assay (ELISA). The results are shown as the means ± SD.

Mentions:
The secretion of these markers analyzed in monocytes supernatant showed similar trend to that observed in plasma. Spontaneous production of cytokines, such as IL-6, TNF-α, Il-1β and VEGF were significantly higher in both groups of cancer patients with and without DVT than those from healthy controls (p<0.0001) (Fig 1). Compared to DVT- cancer patients, DVT+ showed an increased fold change of 1.28 (95% CI: 1.20–1.37; p<0.0001), 1.34 (95% CI: 1.23–1.46; p<0.0001), 1.41 (95% CI: 1.30–1.54; p<0.0001), and 1.61 (95% CI: 1.45–1.78; p<0.0001) for IL-6, TNF-α, Il-1β, and VEGF, respectively. In addition to cytokines, also MMP-9 production and TF activity were higher in DVT+ cancer patients than in those DVT- and in healthy controls (mean ± SD, 103 ± 23, 73 ± 24, 8.8 ± 4, for MMP-9 and mean ± SD, 35.6 ± 6.4, 28 ± 5, 3.3 ± 0.7 for TF activity, respectively). As expected, strong correlations between levels of IL-6, TNF-α, IL-1β, VEGF, MMP-9 and TF in plasma from cancer patients DVT+ and DVT- and those released from monocytes of the same patients were observed (Table 3).

pone.0132496.g001: Cytokines secretion in monocytes from cancer patients with and without DVT.Levels of IL-6, TNF-α, IL-1β, and VEGF were measured in supernatants of purified monocytes from cancer patients with and without DVT by a sensitive enzyme-linked immunosorbent assay (ELISA). The results are shown as the means ± SD.

Mentions:
The secretion of these markers analyzed in monocytes supernatant showed similar trend to that observed in plasma. Spontaneous production of cytokines, such as IL-6, TNF-α, Il-1β and VEGF were significantly higher in both groups of cancer patients with and without DVT than those from healthy controls (p<0.0001) (Fig 1). Compared to DVT- cancer patients, DVT+ showed an increased fold change of 1.28 (95% CI: 1.20–1.37; p<0.0001), 1.34 (95% CI: 1.23–1.46; p<0.0001), 1.41 (95% CI: 1.30–1.54; p<0.0001), and 1.61 (95% CI: 1.45–1.78; p<0.0001) for IL-6, TNF-α, Il-1β, and VEGF, respectively. In addition to cytokines, also MMP-9 production and TF activity were higher in DVT+ cancer patients than in those DVT- and in healthy controls (mean ± SD, 103 ± 23, 73 ± 24, 8.8 ± 4, for MMP-9 and mean ± SD, 35.6 ± 6.4, 28 ± 5, 3.3 ± 0.7 for TF activity, respectively). As expected, strong correlations between levels of IL-6, TNF-α, IL-1β, VEGF, MMP-9 and TF in plasma from cancer patients DVT+ and DVT- and those released from monocytes of the same patients were observed (Table 3).

Bottom Line:
However, their combined role in cancer-associated deep vein thrombosis (DVT) and the molecular mechanisms, involved in its pathophysiology, needs further investigations.Significant correlation between data obtained in plasma and monocyte samples was observed.NF-kB inhibition was associated with decreased levels of all molecules in both cancer DVT+ and DVT-.

Affiliation:
Department of Biomedical and Biotechnological Sciences, Section of General & Clinical Pathology and Oncology, University of Catania, Catania, Italy.

ABSTRACTSeveral studies highlight the role of inflammatory markers in thrombosis as well as in cancer. However, their combined role in cancer-associated deep vein thrombosis (DVT) and the molecular mechanisms, involved in its pathophysiology, needs further investigations. In the present study, C-reactive protein, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1β), matrix metalloproteases-9 (MMP-9), vascular endothelial growth factor (VEGF), tissue factor (TF), fibrinogen and soluble P-selectin, were analyzed in plasma and in monocyte samples from 385 cancer patients, of whom 64 were concomitantly affected by DVT (+). All these markers were higher in cancer patients DVT+ than in those DVT-. Accordingly, significantly higher NF-kB activity was observed in cancer patients DVT+ than DVT-. Significant correlation between data obtained in plasma and monocyte samples was observed. NF-kB inhibition was associated with decreased levels of all molecules in both cancer DVT+ and DVT-. To further demonstrate the involvement of NF-kB activation by the above mentioned molecules, we treated monocyte derived from healthy donors with a pool of sera from cancer patients with and without DVT. These set of experiments further suggest the significant role played by some molecules, regulated by NF-kB, and detected in cancer patients with DVT. Our data support the notion that NF-kB may be considered as a therapeutic target for cancer patients, especially those complicated by DVT. Treatment with NF-kB inhibitors may represent a possible strategy to prevent or reduce the risk of DVT in cancer patients.